Figure 3

Verification of the targeting relationship between miR-153 and Agap2, Bcl2 and Pten. A Base pairings between the mRNA targets (Agap2, Bcl2 and Pten) and miR-153 were predicted by RNAhybrid. B Schematic representation of point mutations. Promoter: PGK, SV40; Fluc: firefly luciferase; Rluc: Renilla luciferase. The electrophoresis results of (C) PCR amplification of the target gene fragment (Agap2 232 bp, Bcl2 376 bp, and Pten 217 bp), D double-enzyme digestion of recombinant pmirGLO-WT vectors, and E double-enzyme digestion of recombinant pmirGLO-MUT vectors. The miR-153 and Agap2 F, Bcl2 G and Pten H mRNA targeting relationships were detected by a dual-luciferase assay system. Calculation formula of the dual-luciferase assay: Ratio = F (experimental group - control group)/R (experimental group–control group). F: Firefly luciferase activity; R: Renilla luciferase activity; The control group: untransfected cells. *P < 0.05, **P < 0.01, and ***P < 0.001 indicate the significance levels of the comparisons between the mimic NC group and the mimic group, as well as between the inhibitor NC group and the inhibitor group.